DQuestion 9An enzyme assay of 2 mL of extract shows 7.3 U/mL and a biuret assay shows 4mg/mL protein on the initial sample. It is salted out and the pellet is washed andresuspended in 0.4 mL buffer. It is re-assayed and shows 12.1 U/mL activity and1.3 mg/mL protein. What is the change in fold purity for this sample? (answer to2 decimal places)

Purification fold of a protein is the specific activity of protein at any particular stage during…

Answered: An enzyme assay of 2 mL of extract…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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(b) Both laboratories used 10 micrograms of protein each in their kinetic assays. Protein concentrations weredetermined by the Bradford protein assay. Assay conditions employed in the two labs (pH, temperature,etc.) were also identical. What would be the most plausible cause for the discrepancy in the Vmax valuesfor the compound I? Explain.Recall that the Bradford assay measures total protein amounts in sample solution based on complexformation between a dye and proteins. Also, the assay solution used in both labs does not contain anyinhibitors.
topic: Bradford AssayThere are numerous methods of protein determination in use, but this module focuses on the Bradford assay.The Bradford assay is a dye-binding method that employs Coomassie Brilliant Blue G-250, whose structureis shown in Figure 2.3.4.1. Coomassie Brilliant Blue G-250 is a dye that interacts with proteins throughhydrophobic and electrostatic interactions. What are the identities and functions of the components of the Bradford reagent in protein contentdetermination?
During incubation, prepare dilutions of the standard antiserum which constitutes the standard curve for the assay (concentration in ng/mL). Add 500 µL of PBS-milk to the supplied microtube containing 500 µL of standard antiserum to obtain Standard 1 [500 ng/mL]. Identify seven microtubes for standards 2 to 7 and place 500 µL of PBS-milk in each. Calculate how much of antiserum is used in each standard ?
Match the types of data to the example. Precise measurements Assay standard Positive control Quantitative results Negative control Qualitative results Standard curve…
5. The data in the table below were collected during a laboratory session assaying for thekinetics of Acid Phosphatase. Using a simple flow diagram, show how you would design theexperiment and focus on the principles of the assay and the major components for theexperimental design. Concentration (mM) Absorbance1 (410nm) Absorbance2 (410nm)0 0.007 0.0081 0.051 0.0672 0.078 0.0965 0.168 0.1558 0.234 0.25210 0.297 0.31620 0.523 0.514 30 0.759 0.719
pls explain Increasing the saturation of the ammonium sulfate is a prerequisite in isolating a target protein that is rich in Cys and Tyr residues. Which of the following techniques should be considered in accurately quantifying the isolated protein?I. Running the isolated protein in a dialysis or GFC set up.II. Using Biuret or BCA assay as the colorimetric quantitation method.III. Using Bradford or Lowry assay as the colorimetric quantitation method.A. I onlyB. II onlyC. I and IIID. I, II and III. Bradford Assay is most suitable to use when the extraction buffer is below the target protein’s pI. This is so because the protein would be morea. Positively charged allowing the CBB G-250 dye to bind via its sulfonate groups.b. Negatively charged allowing the CBB G-250 dye to bind via its sulfonate groups.c. Neutrally charged allowing the CBB G-250 dye to bind via its sulfonate groups.d. Zwitterionic allowing the CBB G-250 dye to bind via its sulfonate groups.
Experiment: Bradford protein assaygive the answers (4-5 lines) of review questions in the end. answer should be logical and understandable and without plagiarism. avoid copy pasting. Background information:The Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly accurate and samples that are out of range can be retested within minutes. The Bradford is recommended for general use, especially for determining protein content of cell fractions and assessing protein concentrations for gel electrophoresis. The method described below is for a 100 µl sample volume using 5 ml color reagent. It is sensitive to about 5 to 200 micrograms protein, depending on the dye quality. In assays using 5 ml color reagent prepared in lab, the sensitive range is closer to 5 to 100 µg protein. Scale down the volume for the "micro assay procedure," which uses 1 ml cuvettes. Protocols, including use of microliter plates are described in the flyer that comes with…
Plz choose all correct option and Do explain. Which of the following descriptions about tandem affinity purification is not correct? a.The interaction between calmodulin-binding peptides and calmodulin beads can be broken by adding EDT b.In the first round of elution, TEV protease cleaves the cleavage site on the TAP-fused target protein. c.It enables the purification of protein complexes composed of multiple protein components. d.It is possible to perform two rounds of purification using a single TAP tag. e.The protein A tag binds to beads that have lgG attached to them.
Calculation for a neutral red assay please include steps since it is essential for the report and it is marked.
1. Suppose a protein sample with a fragment containing the following amino acid sequence is subjected to various chemical assay/tests. - Ala-Gly-Trp-Phe-Met-Cys- What is observed when the protein sample is subjected to Millon’s test? Violet interface Red precipitate/solution Brown/black precipitate Yelllow product No observable result 2. Suppose a protein sample with a fragment containing the following amino acid sequence is subjected to various chemical assay/tests. - Ala-Tyr-Trp-Phe-Cys-Gly - What is observed when the protein sample is subjected to lead sulfide test? Violet interface Red precipitate/solution Brown/black precipitate Yelllow product No observable result
Pls explain If the voltage were NOT increased at the start of the resolving gel run, the overall experiment time will ______, while the calculated electrophoretic mobilities will ______.a. increase, increaseb. increase, decreasec. decrease, increased. decrease, decreasePick all that are TRUE regarding analysis of quaternary structures of proteins using polyacrylamide electrophoresis:I. The added β-mercaptoethanol disrupts S--S bonds bridging the polypeptide chains causing the appearance of higher Rf bands compared to the native protein run. II. Heating up any protein before subjecting to SDS-PAGE will always result in the formation of more than one band.III. A good asymmetrical gel layout would be : (Lane 1) MW ladder, (2) native protein, (3) protein + β-ME, (4) protein + HCL, (5) protein + β-ME + HCl.IV. Formation of a single band in the protein + β-ME + HCl run, whose Rf is lower than the native run, could be indicative that the protein is a homodimer.A. I onlyB. I and IIC. II and…
A protein gives a single band on SDS gel electrophoresis, as shown inlanes 1 and 2 below. There is little, if any, effect from adding β-mercaptoethanol (BME) to the sample; if anything, the protein runs a little bitslower. When treated with the proteolytic enzyme thrombin and electrophoresis in the absence of BME, the protein migrates a bitmore rapidly (lane 3). But if BME is present, two much more rapidlymigrating bands are found (lane 4). Explain these results in terms of amodel for the protein.

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